Reading...
Play button
Play button
Use LEFT and RIGHT arrow keys to navigate between flashcards;
Use UP and DOWN arrow keys to flip the card;
H to show hint;
A reads text to speech;
20 Cards in this Set
- Front
- Back
|
What are the types of tech utilzed in molec genetics?
|
Enzyme, hybridization, amplification, DNA seq techniques
|
|
|
What are restriction enzymes?
|
Endonucleases (called molecular scissors)= cut double stranded DNA w/ seq @ palindromes. Some produce blunt & some sticky ends
|
|
|
What are some clinical applications of restriction enzymes?
|
Southern blot= detects DNA seq, chrom rearrangements, del, repeats & amp. Restriction Fragment Polymprphism (RFLP)= diagnosis diseases w/ point mutations that change restriction sites (beta thal)
|
|
|
What is hybridization?
|
Formation of a stable double-stranded nucleic structure due to complementary base pairing & the formation of H-bonds. Used to detect a nucleic acid seq. Target is hybridized to a probe
|
|
|
What is a probe?
|
A fragment os DNA used to detect specific DNA or RNA molec in a mixture of molec. Is complementary to the target DNA or RNA seq & hybrids to it
|
|
|
What are the labels used to detect probes?
|
*Fluorescence, radioactive, chemiluminescent, Avidin-Biotin peroxidase, Digoxin-Antidigoxin
|
|
|
What are the types of nucleic acid hybridization tech?
|
Southern blot (DNA), Northern blot (RNA), Dot blot, FISH, CGH, Microarrays
|
|
|
What is Southern Blotting?
|
A tech for detecting specific DNA frag in a complex mixture. The target DNA can be a single gene or a part of a larger piece of DNA. Obsolete due to PCR
|
|
|
Describe the process or Southern blotting?
|
Restriction frag-> electrophoresis, blotting-> hybridization w/ probe-> autoradiography
|
|
|
How are some applications of Southern blotting in the clinical lab?
|
Hemophilia A & Fragile X
|
|
|
What causes Hemophilia A?
|
Caused by mutations in the FVIII gene @ Xq28 (most due to inversions)
|
|
|
What is FISH?
|
In situ hybridization of a fluoresently labeled probe to metaphase (dividing) or interphase (nondividing) chrom, whole cells, or tissue sections
|
|
|
How is FISH utilized?
|
Check the presence or absence, copy #, & chrom location of particular DNA seq in chrom of a patient (genetic disease & cancer). Can use multi probes
|
|
|
What is CGH?
|
A general method for looking @ genomic copy # variants. Detects microdeletions or additions. Uses microarrays to examine thousands of probes simultaneously. Don't need to know location of variant. Can scan entire genome
|
|
|
What is PCR?
|
Many applications. Billions of copies can be made from 1 copy of target DNA. Can synthesize unlimited amts. Exponential inc
|
|
|
What is real-time PCR?
|
Quantitative PCR tech that is used extensively in molec microbio
|
|
|
What is DNA seq?
|
Determing the exact nucleotide composition of a DNA fragment
|
|
|
What is Dideoxy method of Seq?
|
Sanger method. Based on synthesis of DNA in vitro in the presence of dideoxynucleotides (chain terminators)
|
|
|
Describe the structure of dideoxynucleotides?
|
Has a hydrogen @ the 3' carbon instead of an OH group. Can form phosphodiester bond w/ the next nucleotide so they terminate DNA elongation. Animation uses Shockwave
|
|
|
What are some applications of DNA seq?
|
BRCA1 &2, alpha & beta thal mutation, MLH1 & 2 (HNPCC)
|
